Role of antimullarian hormone in the diagnosis of sonographically inconclusive polycystic ovary syndrome

Antimullarian hormone (AMH) is found to be a vital tool for the diagnosis of polycystic ovary syndrome (PCOS). AMH may help in the diagnosis of sonographically inconclusive cases of PCOS. This study measured the AMH level in PCOS to assess its impacts on the diagnosis of the syndrome. This cross-sectional study included 160 newly diagnosed females with PCOS who were diagnosed using a modified revised Rotterdam criteria. Fasting blood was collected to measure AMH by enzyme-linked immunosorbent assay and other hormones [total testosterone, luteinizing hormone and follicle-stimulating hormone] were measured by chemiluminescent microparticle immunoassay. Ovarian USG was done in the follicular phase of the menstrual cycle. Serum AMH≥ 3.5 ng/mL and ovarian volume >10 mL was considered as a combined marker of polycystic ovary (PCO). USG-PCO criteria could detect 84.38% PCO, whereas AMH-PCO criteria 67.5%. There was a lack of agreement between USG-PCO and AMH-PCO criteria [κ=-0.004] in PCOS. AMH-PCO criteria identified 68% of PCO patients undiagnosed by USG-PCO criteria [17/25]. Age [β=-0.172, p=0.040], systolic [β =-0.213, p=0.037] and diastolic blood pressure [β=0.301, p=0.004] had significant predictive associations with AMH by linear regression. AMH had a fair discriminating index for combined-PCO [AUC=0.824] in PCOS patients. In conclusion, AMH assessment can help detect PCOS patients who are inconclusively diagnosed by USG-PCO criteria. BSMMU J 2022; 15(2): 65-6

Spectrum of thyroid dysfunctions among hospitalized patients with non-critically ill coronavirus disease 2019: A cross-sectional study

Background: Patients with coronavirus disease 2019 (COVID-19) particularly critically ill ones may present with different types of thyroid abnormalities. However, data regarding thyroid function tests (TFTs) among noncritical patients with COVID-19 are scarce. This study aimed to assess thyroid functions and their associations with the severity of illness among non-critically ill hospitalized patients with COVID-19. Methods: This cross-sectional study assessed TFTs in 87 (aged 18-65 years) RT-PCR-confirmed COVID-19 patients admitted to a tertiary-care hospital in Bangladesh. Diagnosis of non–critical illness and severity (mild, moderate, and severe) were defined by WHO’s interim guidance. Patients having known thyroid dysfunctions or taking drugs that may affect thyroid functions were excluded from the study. Serum TSH, FT4, and FT3 were measured by chemiluminescent immunoassay. Results: Majority of the patients (72%) had normal thyroid function. Among the abnormalities, the highest frequency was isolated hyperthyroxinemia (12.6%) and the rest were subclinical hypothyroidism (6.9%), subclinical thyrotoxicosis (4.6%), thyrotoxicosis (2.3%), isolated tri-iodothyroninemia (1.1%), and hypothyroidism (1.1%).  Serum TSH, FT4, and FT3 levels were similar across the spectrum of noncritical illness. No significant correlation was found between the inflammatory markers (C-reactive protein, ferritin, and D-dimer) and TSH levels. Conclusions: More than one-fourth of non-critically ill hospitalized patients with COVID-19 presented with a spectrum of thyroid abnormalities with isolated hyperthyroxinemia being the most common. However, TFTs had no significant associations with the severity of illness among noncritically ill patients with COVID-19. Bangabandhu Sheikh Mujib Medical University Journal 2023;16(2): 81-86

Short-term and low-dose liraglutide plus metformin decreased body mass index and insulin resistance more than metformin alone in obese women with polycystic ovary syndrome: An open-label randomized controlled study

Background and objectives: Reduction of weight improves different manifestations of polycystic ovary syndrome (PCOS). This study compared the effects of liraglutide plus metformin versus metformin alone on weight loss and metabolic profiles in obese women with PCOS. Methods: This open-label randomized controlled clinical trial consecutively recruited newly-diagnosed PCOS patients of reproductive age with obesity (body mass index ≥ 27.5 kg/m2). Following randomization into two equal groups, Group-1 received treatment with metformin 1000 mg daily alone while Group-2 was given metformin 1000 mg plus subcutaneous (SC) liraglutide 1.2 mg daily for 12 weeks. Anthropometric, biochemical and hormonal data and ovarian morphology were assessed at baseline and after 12 weeks. Clinical information and side effects were recorded every four weeks after initiation of the treatment. Glucose, lipids, and all hormones were analyzed by glucose oxidase, precipitation method, and chemiluminescent microparticle immunoassay respectively. Insulin resistance was measured by homeostatic model assessment (HOMA-IR). Results: Study included 30 participants comprising 15 for each group. Among 15 participants, 5 dropped out from the Group-1 and 1 dropped out from the Group-2. The final analysis was done among 24 participants (Gr-1: 10 and Gr-2: 14). Waist and hip circumference (WC, HC) significantly (p <0.05) decreased in patients treated with only metformin. Menstrual irregularity, BMI (body mass index), HC, systolic blood pressure (BP), 2h-OGTT glucose, fasting insulin, and HOMA-IR significantly (p < 0.05) decreased in the patients of Group-2 after 12 weeks compared to baseline status. Percentage changes of weight, BMI and HOMA-IR improved significantly (p < 0.05) in cases of Group-2 than those in Group-1. Side effects were though numerically higher in the Group-2 patients, but reduced with time. Conclusions: Addition of liraglutide with metformin was superior to metformin alone for lowering of BMI and insulin resistance among obese PCOS women with acceptable side effects. IMC J Med Sci. 2024; 18(1):002. DOI: https://doi.org/10.55010/imcjms.18.002 *Correspondence: Muhammad Abul Hasanat, Room# 1524, Level-15, Block-D, Bangabandhu Sheikh Mujib Medical University (BSMMU), Shahbag, Dhaka-1000, Bangladesh. ORCID iD: orcid.org/0000-0001-8151-9792; Email: [email protected]

Mutation Spectrum in TPO Gene of Bangladeshi Patients with Thyroid Dyshormonogenesis and Analysis of the Effects of Different Mutations on the Structural Features and Functions of TPO Protein through In Silico Approach

Although thyroid dyshormonogenesis (TDH) accounts for 10-20% of congenital hypothyroidism (CH), the molecular etiology of TDH is unknown in Bangladesh. Thyroid peroxidase (TPO) is most frequently associated with TDH and the present study investigated the spectrum of TPO mutations in Bangladeshi patients and analyzed the effects of mutations on TPO protein structure through in silico approach. Sequencing-based analysis of TPO gene revealed four mutations in 36 diagnosed patients with TDH including three nonsynonymous mutations, namely, p.Ala373Ser, p.Ser398Thr, and p.Thr725Pro, and one synonymous mutation p.Pro715Pro. Homology modelling-based analysis of predicted structures of MPO-like domain (TPO142-738) and the full-length TPO protein (TPO1-933) revealed differences between mutant and wild type structures. Molecular docking studies were performed between predicted structures and heme. TPO1-933 predicted structure showed more reliable results in terms of interactions with the heme prosthetic group as the binding energies were -11.5 kcal/mol, -3.2 kcal/mol, -11.5 kcal/mol, and -7.9 kcal/mol for WT, p.Ala373Ser, p.Ser398Thr, and p.Thr725Pro, respectively, implying that p.Ala373Ser and p.Thr725Pro mutations were more damaging than p.Ser398Thr. However, for the TPO142-738 predicted structures, the binding energies were -11.9 kcal/mol, -10.8 kcal/mol, -2.5 kcal/mol, and -5.3 kcal/mol for the wild type protein, mutant proteins with p.Ala373Ser, p.Ser398Thr, and p.Thr725Pro substitutions, respectively. However, when the interactions between the crucial residues including residues His239, Arg396, Glu399, and His494 of TPO protein and heme were taken into consideration using both TPO1-933 and TPO142-738 predicted structures, it appeared that p.Ala373Ser and p.Thr725Pro could affect the interactions more severely than the p.Ser398Thr. Validation of the molecular docking results was performed by computer simulation in terms of quantum mechanics/molecular mechanics (QM/MM) and molecular dynamics (MD) simulation. In conclusion, the substitutions mutations, namely, p.Ala373Ser, p.Ser398Thr, and p.Thr725Pro, had been involved in Bangladeshi patients with TDH and molecular docking-based study revealed that these mutations had damaging effect on the TPO protein activity

Sequenced samples that were used for the HRM method setup.

Sequenced samples that were used for the HRM method setup.</p

Unknown samples used for HRM method validation.

TPO (Thyroid Peroxidase) is known to be one of the major genes involved in congenital hypothyroid patients with thyroid dyshormonogenesis. The present study aims to validate high-resolution melting (HRM) curve analysis as a substitute method for Sanger sequencing, focusing on the frequently observed non-synonymous mutations c.1117G>T, c.1193G>C, and c.2173A>C in the TPO gene in patients from Bangladesh. We enrolled 36 confirmed cases of congenital hypothyroid patients with dyshormonogenesis to establish the HRM method. Blood specimens were collected, and DNA was extracted followed by PCR and Sanger sequencing. Among the 36 specimens, 20 were pre-sequenced, and variants were characterized through Sanger sequencing. Following pre-sequencing, the 20 pre-sequenced specimens underwent real-time PCR-HRM curve analysis to determine the proper HRM condition for separating the three variations from the wild-type state into heterozygous and homozygous states. Furthermore, 16 unknown specimens were subjected to HRM analysis to validate the method. This method demonstrated a sensitivity and specificity of 100 percent in accurately discerning wild-type alleles from both homozygous and heterozygous states of c.1117G>T (23/36; 63.8%), c.1193G>C (30/36; 83.3%), and c.2173A>C (23/36; 63.8%) variants frequently encountered among 36 Bangladeshi patients. The HRM data was found to be similar to the sequencing result, thus confirming the validity of the HRM approach for TPO gene variant detection. In conclusion, HRM-based molecular technique targeting variants c.1117G>T, c.1193G>C, and c.2173A>C could be used as a high throughput, rapid, reliable, and cost-effective screening approach for the detection of all common mutations in TPO gene in Bangladeshi patients with dyshormonogenesis.</div

Differential curves for specimens targeting the c.2173A>C variant in exon 12.

Discernable changes in difference curves were showing specimens with homozygous and heterozygous states were clearly distinguishable from the wild-type, as manifested by the difference in relative fluorescence unit.</p

Difference curves for specimens targeting the c.1193G>C variant in exon 8.

Discernable changes in difference curves were showing that the specimens with homozygous and heterozygous states were clearly distinguishable from the wild-type states, as manifested by the difference in relative fluorescence unit.</p

Normalized melt curves for specimens targeting the c.2173A>C variant in exon 12.

Discernable changes in normalized melt curves were showing that the specimens with homozygous and heterozygous states were clearly distinguishable from the wild-type alleles, as manifested by the difference in relative fluorescence unit.</p

Difference curves generated by specimens targeting the c.1117G>T variant in exon 8.

Discernable changes in three difference curves were showing that the specimens with homozygous and heterozygous states were clearly distinguishable from the wild-type allele, as manifested by the difference in relative fluorescence unit.</p